O-020. Initiation of human primordial follicle growth in vitro in ultrathin slices of ovarian cortex

Picton, Helen M.; Mkandla, A.; Salha, O.; Wynn, PC; Gosden, R. G.

doi:10.1093/humrep/14.suppl_3.11

Cited by 14 publications

(17 citation statements)

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“…In all IVG protocols, it is vital to optimise: i) the supply of nutrients, electrolytes, antioxidants, amino acids, energy substrates, vitamins and growth factors as these components of the culture environment are essential to sustain viability and growth; and ii) the removal of all waste products such as ammonia that may accumulate, as these waste secretions may be detrimental to growth. A variety of base media have been used for IVG of follicles from different species, these include: minimum essential medium (Eppig & Schroeder 1989, Spears et al 1994, Cortvrindt et al 1996, Jewgenow 1998, Newton et al 1999, Picton et al 1999b; Waymouth medium (Eppig & O'Brien 1996, Muruvi et al 2005; and McCoy's 5a medium (Telfer et al 2008). The base media used for follicle culture is supplemented with different culture additives including: antibiotics/antimycotics; commercial preparations of insulin, transferrin and selenium ; physiologically relevant levels of insulin, IGF1 and IGF2 (Newton et al 1999, Picton et al 1999b, Louhio et al 2000; GDF9 (Hreinsson et al 2002); activin (Telfer et al 2008); and 8-bromo cAMP and guanosine monophosphate as apoptosis inhibitors (Scott et al 2004b, Zhang et al 2004.…”

Section: The Follicle Culture Environment In Vitromentioning

confidence: 99%

“…In situ cortical cultures have been conducted for a number of species over varying time frames with or without extracellular matrix and high levels of serum or in serum-free medium (Picton et al 1999b, Webber et al 2007, Telfer et al 2008. During the in situ culture pieces of ovarian cortex are placed onto a physical support, such as a porous membrane insert, in culture wells and the tissue is then covered with a meniscus of culture media (Devine et al 2002).…”

Section: Primordial and Primary Follicle Culturementioning

confidence: 99%

“…Using this approach, follicles may remain viable and are morphologically normal after 1-3 weeks of culture. Using this method, recruitment of primordial follicle growth has been demonstrated in a number of species (cows: Wandji et al 1996, 1997, Braw-Tal & Yossefi 1997, Fortune et al 1998baboons: Fortune et al 1998, hamsters: Yu & Roy 1999, humans: Hovatta et al 1999, Picton et al 1999b. Furthermore, multilayer preantral follicles have been isolated following the culture of fragments of ovarian cortex (Telfer et al 2008) or after culture of whole mouse ovaries (Eppig & O'Brien 1996).…”

Section: Primordial and Primary Follicle Culturementioning

confidence: 99%

“…One of the main reasons for the lack of progress is that it is technically more difficult to recover follicles from these species, as the cortex has a more dense and fibrous consistency (Telfer 1996) and follicle isolation frequently requires the use of aggressive enzyme-based protocols. Nevertheless, in recent years, it has proved possible to maintain the three-dimensional integrity of ruminant (Newton et al 1999, Gutierrez et al 2000, Picton et al 2003) and human follicles (Picton et al 1999b, Telfer et al 2008. The philosophy that underpins this system is that cultured GCs form distinct clumps of rounded cells that closely resemble the morphology (Chang et al 1977) and steroidogenic phentoype of GCs in vivo (Campbell et al 1996, Picton et al 1999a) rather than acquiring a fibroblastic character associated with attachment to the plastic culture dish.…”

Section: Preantral Follicle Culture In Large Domestic Animals and Humansmentioning

confidence: 99%

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The in vitro growth and maturation of follicles

Picton

Harris²,

Muruvi³

et al. 2008

Reproduction

242

190

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The development of technologies to grow oocytes from the most abundant primordial follicles to maturity in vitro holds many attractions for clinical practice, animal production technology and research. The production of fertile oocytes and live offspring has been achieved in mice following the long-term culture of oocytes in primordial follicles from both fresh and cryopreserved ovarian tissue. In contrast, in non-rodent species advances in follicle culture are centred on the growth of isolated preantral follicles. As a functional unit, mammalian preantral follicles are well-suited to culture but primordial and primary follicles do not grow well after isolation from the ovarian stroma. The current challenges for follicle culture are numerous and include: optimisation of culture media and the tailoring of culture environments to match the physiological needs of the cell in vivo; the maintenance of cell-cell communication and signalling during culture; and the evaluation of the epigenetic status, genetic health and fertility of in vitro derived mature oocytes. In large animals and humans, the complete in vitro growth and maturation of oocytes is only likely to be achieved following the development of a multistage strategy that closely mimics the ovary in vivo. In this approach, primordial follicle growth will be initiated in situ by the culture of ovarian cortex. Isolated preantral follicles will then be grown to antral stages before steroidogenic function is induced in the somatic cells. Finally, cytoplasmic and nuclear maturation will be induced in the in vitro derived oocytes with the production of fertile metaphase II gametes.

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Section: The Follicle Culture Environment In Vitromentioning

confidence: 99%

Section: Primordial and Primary Follicle Culturementioning

confidence: 99%

Section: Primordial and Primary Follicle Culturementioning

confidence: 99%

Section: Preantral Follicle Culture In Large Domestic Animals and Humansmentioning

confidence: 99%

See 2 more Smart Citations

The in vitro growth and maturation of follicles

Picton

Harris²,

Muruvi³

et al. 2008

Reproduction

242

190

View full text Add to dashboard Cite

show abstract

“…Most 3D culture systems utilize serum-free media [6,7], alginate gels [8][9][10][11][12] or oocyte encapsulation via collagen [13,14]. These methods adequately provide physical support for follicular growth but fail to recreate endocrine and paracrine interactions between the granulosa and theca cells critical for in vivo follicular maturation.…”

Section: Introductionmentioning

confidence: 99%

In vitro maturation of oocytes via the pre-fabricated self-assembled artificial human ovary

Krotz

Robins

Ferruccio

et al. 2010

J Assist Reprod Genet

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Purpose Create a 3-Dimensional artificial human ovary to mature human oocytes. Methods Theca and granulosa cells were isolated from antral follicles of reproductive-aged women, seeded into micro-molded gels and self-assembled into complex 3D microtissues. Immunohistochemistry and live-dead staining confirmed theca cell identity and cellular viability at one week respectively. Placement of granulosa cell spheroids or cumulus-oocyte complexes into theca cell honeycomb openings resulted in creation of an artificial human ovary. Oocytes from this construct were assessed for polar body extrusion.Results Theca and granulosa cells self-assembled into complex microtissues, remaining viable for one week. At 72 h after artificial human ovary construction, theca cells completely surrounded the granulosa spheroids or COCs without stromal invasion or disruption. Polar body extrusion occurred in one of three COCs assessed. Conclusions An artifical human ovary can be created with self-assembled human theca and granulosa cell microtissues, and used for IVM and future oocyte toxicology studies.

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